Objective To investigate the expression levels of MiR‐96 ,HIF‐1α,Twist and Slug and other predisposing gene in cervical cancer .Methods In this study ,128 patients with cervical cancer treated in our hospital from March 2010 to May 2014 were selected as the observation group ,and 100 cases of healthy people were selected as control group .MiR‐96 ,HIF‐1α,Twist and Slug and other predisposing gene expression levels in cancer tissues were tested .(1) HIF‐1α:HIF‐1α kit was used to detect HIF‐1αmonoclonal antibodies ,the kit was prepared and stained according to the requirements ,and the positive cell rate greater than 10%under the microscope were positive .(2) MiR‐96 ,Twist and Slug:total RNA was extracted according to the instructions ,the RNA was reverse transcribed ,the relative expression values MiR‐96 ,Twist and Slug were detected by quantitative PCR .Results (1) HIF‐1αwas not expressed in normal tissues .And in tumor tissues ,the positive expression rate was much higher than that of normal tissue ,there was a significant difference(P<0 .05);(2)the relative expression values of MiR‐96 in tumor tissue were much greater than that of normal tissue (P<0 .05);(3) the relative expression rate of Twist and Slug gene in the tumor tissue were also much higher than that of normal organizations (P<0 .05) .Conclusion Compared with normal tissues ,MiR‐96 ,HIF‐1α,Twist and Slug gene expression in tumor tissues are significantly greater .

Lipopolysaccharide(LPS) can induce cell inflammation through interacting with TLR4. Recent studies have revealed that MD-2 participate in the process of LPS induced signal transduction pathway by forming a complex with TLR4. After binding to the MD-2 of the TLR4/MD-2 complex, LPS can induce TLR4- oligomerization and activate the downstream signal pathway. After being synthesized, most MD-2 can bind to TLR4 at the endoplasmic reticulum /Golgi apparatus and expresse as TLR4/MD-2 complex at the cellular surface. Therefore MD-2 not only can regulate the distribution of TLR4 in the cytoplasm, but also help TLR4 to recognize LPS. Another part of MD-2 can be released into plasma as soluble MD-2(sMD-2). With the help of CD14, sMD-2 would interact with LPS in the plasma to constitute LPS-sMD-2 complex, helping cell who express only TLR4, to recognize LPS, however excessive expressed sMD-2 would repress the LPS signal transduction pathway. In conclusion, MD-2 plays a crucially modulating role in the process of TLR4 mediated endotoxin recognition and signal transduction.

TLR4-MD-2 complex plays a key role in LPS recognition and its signal transduction. These steps are the vital elements of the host's defensive reaction. Studying the functional domain of TLR4 and MD-2 is very important to further understand the mechanism of LPS signal transduction. It was studied the interaction domain of TLR4 and MD-2 in living cells based on gene mutation, gene transfection and fluorescence resonance energy tramsfer(FRET) which is considered as one of the best methods used for intracellular protein-protein interaction study. CY-15P which was fused by CFP and YFP through 15 neutral amino acids was used as positive control, while co-expressed CFP and YFP proteins were used as negative control. The results showed that the ability of TLR4 binding to MD-2 decreased dramatically after the deletion of Glu~(24) ～ Met~(41) in N terminal of TLR4. Aggregation of TLR4 to LPS stimulation was observed, however, TLR4 without the Glu~(24)～ Met~(41) mutation did not aggregate. All these results indicated that TLR4 Glu~(24)～ Met~(41) might be the interaction domain of TLR4 binding to MD-2 and participate in the aggregation effect of TLR4 upon LPS stimulation.

Objective To evaluate the clinical value of double-balloon enteroscopy( DBE)in as-sessing the effect of mucosal healing in patients with moderate small bowel Crohn's disease( CD)treated with low-dose azathioprine. Methods CD patients who were naive to any immunomodulators or biological a-gents with lesions mainly located in ileu were screened by multislice CT enterography and anal-route DBE at baseline. Lesions at 150 cm proximal to ileocecal valve were assessed by DBE with Simple Endoscopic Score for CD( SES-CD)after 12 and 24 months of low-dose azathioprine treatment,respectively. Results A total of 36 patients were enrolled and the average tolerated dose of azathioprine was(61. 8 ± 17. 2)mg/day. The total rates of complete,near-complete,partial and no mucosal healing in 36 patients were 19. 4%(7/36), 5. 6%(2/36),27. 8%(10/36),and 47. 2%(17/36)at month 12 and 30. 6%(11/36),25. 0%(9/36), 33. 3%(12/36),and 11. 1%(4/36)at month 24,respectively. The baseline SES-CD score(OR=2. 71, 95%CI:1. 11-6. 63,P=0. 029)and duration of disease(OR=1. 27,95%CI:1. 10-1. 47,P =0. 001) were two relevant factors associated with mucosal healing of small bowel CD. Conclusion DBE has a signif-icant advantage in assessing post-therapy mucosal healing for patients with small bowel CD. The optimal time point for the first follow-up by DBE is at least 12 months after low-dose azathioprine treatment.

Objective To analyze the efficacy and tolerance of methotrexate(MTX)in remission maintenance of Crohn′s disease (CD).Methods From June 2012 to August 2015 ,49 CD patients who received MTX as mainly treatment medication to maintain remission were enrolled.The pre-medication history,efficacy,dosage and side effects of MTX were analyzed.The effects of inducing strategy on disease recurrence were analyzed.Chi-square test and t test were used for statistical analysis.Results Among the 49 patients,34 (69.4%)received steroids for remission inducing,nine (18.4%)received infliximab for remission inducing and six (12.2%)achieved remission after operation.In the 44 patients treated with azathioprine (AZA)before,the median treatment time was one month and the dosage for withdrawal of AZA was (42.0 ± 14.8)mg/d.The most common reason was leucopenia (81 .8%, 36/44).Till the time point of follow-up,46 of the 49 CD patients still took MTX orally with a median treatment time of 16 months,and the weekly dosage was (12.7 ±2.0)mg.Thirty-one cases (67.4%) achieved clinical stability,while 15 cases (32.6%)underwent clinical recurrence.The median Crohn′s disease activity index (CDAI)was 123.5 ± 66.6.The weekly dosage of clinical stability group was (12.5 ±2.1)mg,and that of clinical recurrence group was (13.0 ±1 .7 )mg,there was no statistically significant difference between the two groups (t =0.802,P =0.426 ).The recurrence rate of steroids-induced remission group was 41 .2% (14/34 ),which was higher than that of infliximab and surgery-induced remission group (1/15),and the differnce was statistically significant (χ2 =5 .177,P =0.023 ). The common side effects were gastrointestinal reaction (26.5 %, 13/49 ), impaired liver function (20.4%,10/49)and leukopenia (12.2%,6/49).Only three cases could not tolerate the side effects and underwent medication withdrawal.Conclusions As a second-line immunosuppressant for maintanence remission in CD,MTX is effective and well-tolerated in patients.So it can be an important option during the long course of CD.

Objective:To compare the osteogenesis ability between human umbilical cord Wharton's Jelly-derived mesenchymal stem cells(hUCWJMSCs) and human periodontal ligament mesenchymal stem cells (hPDLSCs) in vitro.Methods:hUCWJMSCs and hPDLSCs were in vitro cultured.The cell proliferation capacity was examined by MTT assay.After osteogenesis induction culture,ALP activity of the cells was determined,minerialization was observed by alizarin red staining,OPN and Runx2 mRNA expression was analyzed by Real-time PCR.Results:hUCWJMSCs grew faster than hPDLSCs.After osteogenic differentiation induction,hPDLSCs group showed higher ALP level,more mineralized nodule formation and higher Runx2 expression compared with hUCWJMSCs group (P ＜ 0.05);while the OPN expressed higher in hUCWJMSCs than in hPDLSCs (P ＜ 0.05).Conclusion:hUCWJMSCs and hPDLSCs have osteogenesis differentiation potential,hPDLSCs are more osteogenetic.

Objective:To evaluate the effect of dexmedetomidine on pyroptosis in mice with acute renal injury induced by endotoxin and the relationship with miRNA-223-3p.Methods:Thirty-two clean-grade healthy male ICR mice, aged 8-12 weeks, weighing 20-25 g, were divided into 4 groups ( n=8 each) using the random number table method: control group (group C), lipopolysaccharide (LPS) group (group L), LPS plus dexmedetomidine group (group LD), and LPS plus dexmedetomidine plus atipamezole group (group LDT). The model of acute renal injury induced by endotoxin was established by intraperitoneal injection of LPS 400 μg/kg, followed by intraperitoneal injection of LPS 10 mg/kg 8 h later.Dexmedetomidine 40 μg/kg was intraperitoneally injected once every 2 h for 3 times in total starting from 30 min after establishing the model in group LD.Atipamezole 750 μg/kg was intraperitoneally injected immediately after establishing the model, and 30 min later dexmedetomidine 40 μg/kg was intraperitoneally injected once every 2 h for 3 times in total in group LDT.The equal volume of normal saline was intraperitoneally injected in group C. Blood samples were collected from the heart at 24 h after establishing the model, and serum creatinine (Cr) and blood urea nitrogen (BUN) concentrations were measured with an automatic biochemical analyzer.The animals were sacrificed and the left kidney tissues were obtained for microscopic examination of pathological changes after HE staining (with a light microscope) and for determination of the expression of caspase-1 p20, NOD-like receptor thermoprotein structural domain-related protein 3 (NLRP3) and ASC protein and mRNA (by quantitative real-time polymerase chain reaction and Western blot), contents of interleukin-1beta (IL-1β) and IL-18 (by enzyme-linked immunosorbent assay), and rate of pyroptosis in renal cortical cells (by TUNEL). Results:Compared with group C, the concentrations of serum Cr and BUN were significantly increased, the expression of NLRP3, caspase-1 p20 and ASC protein and mRNA in the renal tissues was up-regulated, the contents of IL-1β and IL-18 were increased, the rate of pyroptosis in renal cortical cells was increased ( P<0.05), no significant change was found in the expression of miRNA-223-3p ( P>0.05), and pathological changes of kidney were accentuated in group L. Compared with group L, the concentrations of serum Cr and BUN were significantly decreased, the expression of NLRP3, caspase-1 p20 and ASC protein and mRNA in the renal tissues was down-regulated, the contents of IL-1β and IL-18 were decreased, the rate of pyroptosis in renal cortical cells was decreased, the expression of miRNA-223-3p was up-regulated ( P<0.05), and pathological changes of kidney were attenuated in group LD.Compared with group LD, the concentrations of serum Cr and BUN were significantly increased, the contents of IL-1β and IL-18 were increased, the expression of NLRP3, caspase-1 p20 and ASC protein and mRNA in the renal tissues was up-regulated, the rate of pyroptosis in renal cortical cells was increased, the expression of miRNA-223-3p was down-regulated ( P<0.05), and the pathological changes of kidney were accentuated in group LDT. Conclusion:The mechanism by which dexmedetomidine reduces acute renal injury may be related to up-regulating the expression of miRNA-223-3p and inhibiting pyroptosis in mice.

Objective:To evaluate the relationship between Sestrin2 and mitochondrial DNA (mtDNA)-NOD-like receptor associated protein 3 (NLRP3) inflammasome pathway during endotoxin-induced myocardial injury in mice.Methods:One hundred and eighty-four clean-grade healthy male ICR mice, aged 8-12 weeks, weighing 20-25 g, were used in this study. One hundred and sixty-eight mice were divided into 7 groups ( n=24 each) using the random number table method: normal control group (N group), lipopolysaccaride(LPS) group (L group), mtDNA group, LPS+ mtDNA group (M group), normal control+ negative control adeno-associated virus (AAV-NC)group (NC group), LPS+ mtDNA+ AAV-NC group (MC group), and LPS+ mtDNA+ Sestrin2 overexpression adeno-associated virus (AAV-Sestrin2) group (MSgroup). Another 10 mice were used to detect the transfection effect of AAV-Sestrin2, and the left 6 mice were used for mtDNA extraction. The model of endotoxemia was developed by intraperitoneal injection of LPS 10 mg/kg. mtDNA 5 mg/kg was intraperitoneally injected in mtDNA group, and mtDNA 5 mg/kg was intraperitoneally injected at 30 min after LPS injection in M group.AAV-Sestrin2 150 μl was injected via the tail vein in MS group, and the equal volume of AAV-NC was injected via the tail vein in MC and NC groups. Four weeks after virus injection, LPS 10 mg/kg was intraperitoneally injected and 30 min later mtDNA 5 mg/kg was intraperitoneally injected in MS and MC groups. Blood samples were collected at 24 h after LPS injection for determination of serum creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH) activities (by biochemical assay), concentrations of serum cardiac troponin I (cTnI), interleukin-18 (IL-18) and interleukin-1beta (IL-1β)(by enzyme-linked immunesorbent assay), and expression of mtDNA (by quantitative real-time polymerase chain reaction). The animals were sacrificed after the end of blood sampling and myocardial tissues were obtained for determination of the contents of reactive oxygen species (ROS), total antioxidant capacity (T-AOC), and adenosine triphosphate (ATP) and expression of NOD-like receptor associated protein 3 (NLRP3), active subunit p20 of caspase-1 (caspase-1p20) and apoptosis-associated microprotein (ASC) in myocardial tissues (by Western blot) and for microscopic examination of the pathological changes after HE staining (with a light microscope). Results:Compared with N group, the levels of CK-MB, LDH, cTnI, IL-1β and IL-18 in serum were significantly increased, the expression of mtDNA was up-regulated, the ROS content in myocardial tissues was increased, the T-AOC and ATP contents in myocardial tissues were decreased, the expression of NLRP3, caspase-1p20 and ASC in the myocardial tissues was up-regulated( P<0.05), and the pathological changes of myocardial tissues were aggravated in L group and mtDNA group.Compared with L group and mtDNA group, the levels of CK-MB, LDH, cTnI, IL-1β and IL-18 in serum were significantly increased, the expression of mtDNA was up-regulated, the ROS content in myocardial tissues was increased, the T-AOC and ATP contents in myocardial tissues were decreased, the expression of NLRP3, caspase-1p20 and ASC in the myocardial tissues was up-regulated( P<0.05), and the pathological changes of myocardial tissues were aggravated in M group. Compared with M group, the levels of CK-MB, LDH, cTnI, IL-1β and IL-18 in serum were significantly decreased, the expression of mtDNA was down-regulated, the ROS content in myocardial tissues was decreased, the T-AOC and ATP contents in myocardial tissues were increased, the expression of NLRP3, caspase-1p20 and ASC in the myocardial tissues was down-regulated( P<0.05), and the pathological changes of myocardial tissues were significantly attenuated in MS group. Conclusions:Sestrin2 can reduce endotoxin-induced myocardial injury in mice by alleviating mitochondrial damage, inhibiting oxidative stress, protecting mtDNA from oxidative damage, and then inhibiting mtDNA-NLRP3 inflammasome pathway.

Echinococcus granulosus is the causative agent of cystic echinococcosis with medical and veterinary importance in China. Our main objective was to discuss the genotypes and genetic diversity of E. granulosus present in domestic animals and humans in western China. A total of 45 hydatid cyst samples were collected from sheep, humans, and a yak and subjected to an analysis of the sequences of mitochondrial cytochrome b (cytb) gene. The amplified PCR product for all samples was a 1,068 bp band. The phylogenetic analysis showed that all 45 samples were identified as E. granulosus (genotype G1). Ten haplotypes were detected among the samples, with the main haplotype being H1. The haplotype diversity was 0.626, while the nucleotide diversity was 0.001. These results suggested that genetic diversity was low among our samples collected from the west of China based on cytb gene analysis. These findings may provide more information on molecular characteristics of E. granulosus from this Chinese region.
Animals ; Animals, Domestic/parasitology ; Base Composition ; Base Sequence ; Cattle/*parasitology ; China ; Cytochromes b/*genetics ; DNA, Helminth/genetics ; Echinococcosis ; Echinococcus granulosus/classification/*genetics ; Genetic Variation ; Haplotypes/genetics ; Humans ; Mitochondria/genetics ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction/*veterinary ; Sequence Alignment ; Sequence Analysis, DNA ; Sheep/*parasitology ; Tibet

Objective:To explore the curative effect of surgical treatment for Crohn′s disease (CD), to investigate the timing of surgical intervention and the choice of surgical methods.Methods:From January 1, 2016 to August 31, 2020, at Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, the clinical data of 123 patients with CD and receiving surgical treatment were retrospectively analyzed, which included the type of lesion, the location of lesion, clinical manifestation, surgical method, preoperative inflammatory and nutritional indicators, postoperative recovery of digestive tract function, and the development and treatment of postoperative complications. CD was diagnosed according to Consensus opinion on diagnosis and treatment of inflammatory bowel disease ( Beijing 2018). Patient was classitied according to the Montreal Classification. Postoperative complications were graded according to the Clavien-Dindo Criteria. Mann-Whitney U test was used for statistical analysis. Results:Among 123 patients, according to the Montreal classification, two cases (1.6%) were diagnosed at ≤16 years old (type A1), 66 cases (53.7%) were diagnosed at 17 to 40 years old (type A2), and 55 cases (44.7%) were diagnosed at >40 years old (type A3). The lesions were 52 cases (42.3%) of terminal ileum (L1) type, 20 cases (16.3%) of colon (L2) type, and 51 cases (41.5%) of ileocolon (L3) type. Four cases (3.2%) were non-stenosis and non-penetrating (B1) type, 87 cases (70.7%) were stenosis (B2) type, and 32 cases (26.0%) were penetrating (B3) type. Eighteen patients (14.6%) underwent emergency surgery due to complete intestinal obstruction (10 cases), gastrointestinal perforation (five cases), gastrointestinal bleeding (two cases), and rectovesical fistula complicated with septic shock (one case). One hundred and five patients (85.4%) received selective surgery due to poor conservative treatment effects. 51 cases (41.5%) underwent traditional open surgery and 72 cases (58.5%) underwent laparoscopic surgery. Nineteen patients (15.4%) received temporary or permanent ostomy. The preoperative C reactive protein level of patients with emergency surgery was higher than that of patients undergoing selective surgery ((39.23±24.13) mg/L vs. (11.48±2.68) mg/L), while the levels of plasma albumin (ALB) and pre-ALB were lower than those of patients receiving selective surgery ((29.90±10.60) g/L vs. (38.38±8.30) g/L, (146.00±125.49) mg/L vs. (209.06±61.19) mg/L), and the differences were statistically significant ( Z=9.603, 8.754 and 7.111, all P<0.01). During the follow-up, a total of 23 cases (18.7%) developed postoperative complications, including one case of postoperative intra-abdominal hemorrhage and underwent re-operation (Clavien-Dindo grade Ⅲ complication); four cases of anastomotic leakage after operation; six cases of postoperative paralytic ileus; 11 cases of surgical site infection, all of which were Clavien-Dindo grade Ⅱ complications, and one case of deep venous thrombosis of lower extremity. No patient with severe intraoperative complication was observed, and no patients died during the operation or hospitalization. The postoperative exhaust time of patients was (3.2±1.4) d, the time of open fluid diet was (5.8±0.8) d, the length of hospital stay was (18.0±14.1) d, and the length of postoperative hospital stay was (11.2±8.8) d. Conclusions:The concept of multidisciplinary collaboration should be emphasized in the treatment of CD. Surgical treatment can effectively control the complications and improve the quality of life of patients, but the timing of operation and the choice of surgical methods should be decided prudently after perioperative treatment, multi-disciplinary participated and regulation of the internal environment. The standardized and targeted treatments for the surgical difficulties of inflammatory bowel disease should be conducted.